Combination therapy using an anti-fucosyl-gm1 antibody and an anti-cd137 antibody

ABSTRACT

This disclosure provides combination therapy for treating a subject, such as a subject afflicted with a lung cancer, comprising administering to the subject an anti-fucosyl-GM1 antibody and an anti-CD137 antibody, or antigen-binding portions of either or both.

FIELD OF THE INVENTION

The present invention relates to improved methods of treating cancerusing a combination of therapeutic monoclonal antibodies.

BACKGROUND OF THE INVENTION

Fucosyl-GM1 is a sphingolipid monosialoganglioside composed of aceramide lipid component, which anchors the molecule in the cellmembrane, and a carbohydrate component that is exposed at the cellsurface. Carbohydrate antigens are the most abundantly expressedantigens on the cell surface of cancers (Feizi T. (1985) Nature314:53-7). In some tumor types, such as small cell lung cancer (SCLC),initial responses to chemotherapy are impressive, but chemo-refractoryrelapses rapidly follows. Intervention with novel immunotherapeutics maysucceed in overcoming drug resistant relapse (Johnson D H. (1995) LungCancer 12 Suppl 3:S71-5). Several carbohydrate antigens, such asgangliosides GD3 and GD2, have been shown to function as effectivetargets for passive immunotherapy with mAbs (Irie R F and Morton D L.(1986) PNAS 83:8694-8698; Houghton A N et al. (1985) PNAS 82:1242-1246).Ganglioside antigens have also been demonstrated to be effective targetsfor active immunotherapy with vaccines in clinical trials (Krug L M etal. (2004) Clinical Cancer Research 10:6094-6100; Dickler M N et al.(1999) Clinical Cancer Research 5:2773-2779; Livingston P O et al.(1994) J Clin. Oncol. 12:1036-44). Indeed, serum derived from SCLCpatients who developed antibody titers to fucosyl-GM1 followingvaccination with KLH conjugated antigen, demonstrated specific bindingto tumor cells and tumor specific complement dependent cytotoxicity(CDC). Anti-fucosyl-GM1 titer associated toxicities were mild andtransient and three patients with limited-stage SCLC were relapse-freeat 18, 24, and 30 months (Krug et al., supra; Dickler et al., supra).

Fucosyl-GM1 expression has been shown in a high percentage of SCLC casesand unlike other ganglioside antigens, fucosyl-GM1 has little or noexpression in normal tissues (Nilsson et al. (1984) Glycoconjugate J.1:43-9; Krug et al., supra; Brezicka et al. (1989) Cancer Res.49:1300-5; Zhangyi et al. (1997) Int. J. Cancer 73:42-49; Brezicka etal. (2000) Lung Cancer 28:29-36; Fredman et al. (1986) Biochim. Biophys.Acta 875: 316-23; Brezicka et al. (1991) APMIS 99:797-802; Nilsson etal. (1986) Cancer Res. 46:1403-7). The presence of fucosyl-GM1 has beendemonstrated in culture media from SCLC cell lines, in tumor extractsand serum of nude mouse xenografts and in the serum of SCLC patientswith extensive-stage disease (Vangsted et al. (1991) Cancer Res.51:2879-84; Vangsted et al. (1994) Cancer Detect. Prev. 18:221-9).Fucosyl-GM1 expression has also been observed in a significant fractionof non-small cell lung cancer (NSCLC) samples. WO 07/067992. Thesereports provide convincing evidence for fucosyl-GM1 as a highly specifictumor antigen, which may be targeted by an immunotherapeutic.

An antibody that recognizes fucosyl-GM1 on cancer cells and directstheir destruction, anti-fucosyl GM1 mAb BMS-986012, has entered clinicaltrials for the treatment of subjects with relapsed/refractory small celllung cancer (NCT02247349). See Molckovsky & Siu (2008) J. Hematol.Oncol. 1:20. BMS-986012 is a non-fucosylated antibody and thus exhibitsenhanced ADCC compared to an antibody with typical mammalianglycosylation. Although effective as a single agent, there exists a needfor even more effective cancer therapy based on targeting of fucosylGM1-expressing cancer cells.

SUMMARY OF THE INVENTION

The present invention provides methods for treating a subject afflictedwith cancer, such as a lung cancer, including small cell lung cancer(SCLC), comprising administering to the subject a therapeuticallyeffective combination of agents, such as monoclonal antibodies or anantigen-binding portions thereof, that specifically bind to fucosyl-GM1and CD137. In one embodiment the anti-fucosyl-GM1 antibody ishypofucosylated or non-fucosylated.

In some embodiments, the anti-fucosyl-GM1 mAb competes with BMS-986012,comprises the same CDRs as BMS-986012, comprises the same heavy andlight chain variable domains as BMS-986012, comprises the same heavy andlight chains as BMS-986012, is BMS-986012, or is an antibody drugconjugate of BMS-986012. In one embodiment, the anti-CD137 mAb competeswith BMS-663513, comprises the same CDRs as BMS-663513, comprises thesame heavy and light chain variable domains as BMS-663513, or isBMS-663513.

In one aspect, the invention provides methods of combination therapywith anti-fucosyl GM1 and anti-CD137 agents comprising administration ofthe anti-fucosyl GM1 agent, such as BMS-986012, concurrently withadministration of the anti-CD137 agent.

In a distinct aspect, the invention provides methods of combinationtherapy with anti-fucosyl GM1 and anti-CD137 agents comprisingadministration of the anti-fucosyl GM1 agent, such as BMS-986012, priorto, rather than concurrently with, the anti-CD137 agent, such asBMS-663513. In various embodiments, the anti-fucosyl GM1 agent is firstadministered 0.5, 1, 2, 4, and 7 days prior to first administration ofthe anti-CD137 agent. In various other embodiments, the anti-fucosyl GM1agent is administered at days 1, 22, 43, 64 and 85 and the anti-CD137agent is administered at days 2, 23, 44, 65 and 86.

In some embodiments, the combination therapy of the present invention isadministered to a subject (e.g. a human subject) who suffers from smallcell lung cancer (SCLC). In one embodiment, the subject has previouslyreceived an initial anti-cancer therapy. In another embodiment, the lungcancer is an advanced, metastatic, relapsed, and/or refractory lungcancer.

In certain embodiments, the anti-fucosyl-GM1 mAb is administered at adose ranging from 10 to 2000 mg once every 1, 2, 3 or 4 weeks. In oneembodiment, the anti-fucosyl-GM1 mAb is administered at a dose rangingfrom 400 to 1000 mg once every 3 weeks, with administration ofanti-CD137 mAb at a dose ranging from 1 to 10 mg, e.g. 3 to 8 mg, oneday after each dose of anti-fucosyl-GM1 mAb. In various embodiments, thedose of the anti-fucosyl GM1 mAb is 2-, 10-, 30- or 1000-fold higherthan the dose of anti-CD137 mAb on a weight basis.

In various embodiments, the method comprises one, two, three or fourtreatments, or is continued for as long as clinical benefit is observedor until unmanageable toxicity or disease progression occurs.

In one embodiment, one or both of the antibodies are formulated forintravenous administration. The efficacy of the treatment methodsprovided herein can be assessed using any suitable means, such asreduction in size of the cancer, reduction in number of metastaticlesions over time, complete response, partial response, and stabledisease.

Other features and advantages of the instant invention will be apparentfrom the following detailed description and examples, which should notbe construed as limiting. The contents of all cited references,including scientific articles, newspaper reports, GenBank entries,patents and patent applications cited throughout this application areexpressly incorporated herein by reference in their entireties.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows tumor growth in the mouse DMS79 tumor model as a functionof treatment with anti-fucosyl GM1 BMS-986012 and/or anti-muCD137antibodies. Median tumor volumes are presented for groups of 8 mice ateach data point. See Example 1.

FIG. 2 shows tumor growth in the mouse DMS79 tumor model as a functionof treatment with anti-fucosyl GM1 BMS-986012 and/or cisplatin. Mediantumor volumes are presented for groups of 8 mice at each data point. SeeExample 2.

FIG. 3 shows tumor growth in the mouse DMS79 tumor model as a functionof treatment with anti-fucosyl GM1 BMS-986012 and/or etoposide. Mediantumor volumes are presented for groups of 8 mice at each data point.Combination therapy is significantly more effective than eithermonotherapy. See Example 3.

DETAILED DESCRIPTION OF THE INVENTION

BMS-986012 is a first-in-class fully human monoclonal antibody (mAb)that specifically binds to the fucosyl-GM1 (fuc-GM1) ganglioside.BMS-986012 exhibits high-affinity and dose-dependent saturable bindingto fuc-GM1 and shows no detectable antigen-specific binding to closelyrelated molecule GM1. Because fucosyl GM1 is preferentially found on thesurface of lung cancer cells, BMS-986012 is particularly well suited totreating lung cancer, such as small cell lung cancer (SCLC). BMS-986012,as used herein, always refers to a non-fucosylated antibody whether ornot that is expressly stated.

BMS-986012 is non-fucosylated (lacking fucosylation on the Fc domain).The absence of the fucosyl group in BMS-986012 confers higher affinityfor Fc receptors resulting in enhanced antibody-dependent cellularcytotoxicity (ADCC). Furthermore, the antibody was shown to mediatepotent complement dependent cytotoxicity (CDC) as well asantibody-dependent cellular phagocytosis (ADCP). See, e.g., WO2007/067992, the content of which is expressly incorporated herein byreference in its entirety.

Although BMS-986012 is effective as monotherapy in treatment of lungcancer, improved methods of treatment are always desired.

CD137 (4-1BB) is a T cell co-stimulatory receptor induced on TCRactivation. Nam et al. (2005) Curr. Cancer Drug Targets 5:357; Watts etal. (2005) Ann. Rev. Immunol. 23:23. In addition to its expression onactivated CD4⁺ and CD8⁺ T cells, CD137 is also expressed on CD4⁺ CD25⁺regulatory T cells, activated natural killer (NK) and NK-T cells,monocytes, neutrophils, and dendritic cells. Its natural ligand, CD137L,has been described on antigen-presenting cells including B cells,monocyte/macrophages, and dendritic cells. Watts et al. (2005) Ann. Rev.Immunol. 23:2. On interaction with its ligand, CD137 leads to increasedTCR-induced T-cell proliferation, cytokine production, functionalmaturation, and prolonged CD8⁺ T-cell survival. Nam et al. (2005) Curr.Cancer Drug Targets 5:357; Watts et al. (2005) Ann. Rev. Immunol. 23:23.Use of antibodies to activate the CD137 pathway for treating cancer hasentered clinical trials. Li et al. (2013) Clin. Pharmacol. 5(Suppl1):47; Sznol et al. (2008) J Clin. Oncol. 26(Suppl. 15):3007.

Applicants observed that administration of anti-fucosyl GM1 antibody toa mixture of DMS-79 (fucosyl GM1-expressing) tumor cells and NK cells invitro induced a dose-dependent enhancement in CD137 expression on the NKcells, increasing the percentage of CD137+NK cells from less than 4% toover 35% at the highest doses (data not shown). See also Kohrt et al.(2011) Blood 117:2423 (disclosing a similar effect with anti-CD20); Linet al. (2008) Blood 112:699. Without intending to be limited by theory,it is possible that BMS-986012 bound to fucosyl GM1 on tumor cells alsobinds to Fc receptors on NK cells, and that this interaction with the NKcell Fc receptor is what triggers increased CD137 expression. BecauseCD137 is an activator of NK cells, CD137 agonism might be helpful inenhancing NK cell-mediated anti-tumor effects. See U.S. Pat. No.7,288,638; Melero et al. (2007) Nat. Rev. Cancer 7:95; Takeda et al.(2010) J. Immunol. 184:5493; Murillo et al. (2008) Clin. Cancer Res.14:6895. Combination therapy methods of present invention are based onincreasing CD137 signaling, for example using an agonist anti-CD137antibody, and the enhancement of this effect by prior or concurrentadministration of anti-fucosyl GM1 antibodies to increase CD137expression on NK cells, thus maximizing the effect of anti-CD137therapy. Equivalently, the invention may be viewed as augmenting theanti-tumor (cell killing) effect of anti-fucosyl GM1 antibody byadministering an agonist anti-CD137 antibody that enhances the NKcell-mediated anti-tumor effects.

The specific combination of anti-fucosyl GM1 antibody with anti-CD137antibody is particularly beneficial because treatment with anti-fucosylGM1 antibody enhances CD137 expression on NK cells, maximizing theeffect of the agonist anti-CD137 antibody. One benefit of suchcombination therapy is that it may preferentially direct the effects ofagonist anti-CD137 therapy toward the desired target cells (fucosylGM1-expressing lung cancer cells) by enhancing their CD137 expression,and away from general systemic NK cell stimulation that may give rise toimmune-mediated side effects. Kohrt et al. (2011) Blood 117:2423. Suchtargeting may allow for lower dosing of agonist CD137 antibodies, thusminimizing undesired toxicities.

In some embodiments of the present invention, the anti-fucosyl GM1antibody is administered prior to administration of anti-CD137 antibody.Without intending to be limited by theory, such pre-treatment withanti-fucosyl GM1 antibody allows binding to fucosyl GM1 on target tumorcells, ligation of the Fc region of the anti-fucosyl GM1 antibody to Fcreceptors on NK cells, and most importantly upregulation of CD137 on NKcells, prior to administration of anti-CD137. As a consequence, at the(subsequent) time of dosing with anti-CD137 there exists in the subjecta pre-existing population of NK cells in proximity to the target tumorcells and with elevated CD137 expression, such that binding (andeffects) of anti-CD137 will be preferentially directed to the targettumor cells.

I. Definitions

In order that the present disclosure may be more readily understood,certain terms are first defined. As used in this application, except asotherwise expressly provided herein, each of the following terms shallhave the meaning set forth below. Additional definitions are set forththroughout the application.

“BMS-986012,” as used herein, refers to a non-fucosylated anti-fucosylGM1 mAb comprising heavy chains of SEQ ID NO: 3 and light chains of SEQID NO: 4. BMS-986012 also comprises heavy chain variable domains of SEQID NO: 1 and light chain variable domains of SEQ ID NO: 2. BMS-986012also comprises CDR sequences of SEQ ID NO: 5 (CDRH1), SEQ ID NO: 6(CDRH2), SEQ ID NO: 7 (CDRH3), SEQ ID NO: 8 (CDRL1), SEQ ID NO: 9(CDRL2), SEQ ID NO: 10 (CDRL3).

“BMS-6633513,” as used herein, refers to an anti-CD137 mAb comprisingheavy chains of SEQ ID NO: 13 and light chains of SEQ ID NO: 14, alsoknown as urelumab. BMS-6633513 also comprises heavy chain variabledomains of SEQ ID NO: 11 and light chain variable domains of SEQ ID NO:12. BMS-6633513 also comprises CDR sequences of SEQ ID NO: 15 (CDRH1),SEQ ID NO: 16 (CDRH2), SEQ ID NO: 17 (CDRH3), SEQ ID NO: 18 (CDRL1), SEQID NO: 19 (CDRL2), SEQ ID NO: 20 (CDRL3).

Target proteins referenced herein (also referred to as antigens), suchas CD137, are intended to refer to their human orthologs (e.g. huCD137;NP_001552; GeneID 3604) unless otherwise indicated or clear from thecontext.

“Administering” refers to the physical introduction of a compositioncomprising a therapeutic agent to a subject, using any of the variousmethods and delivery systems known to those skilled in the art.Preferred routes of administration for the anti-fucosyl-GM1 antibodyinclude intravenous, intramuscular, subcutaneous, intraperitoneal,spinal or other parenteral routes of administration, for example byinjection or infusion. The phrase “parenteral administration” as usedherein means modes of administration other than enteral and topicaladministration, usually by injection, and includes, without limitation,intravenous, intramuscular, intraarterial, intrathecal, intralymphatic,intralesional, intracapsular, intraorbital, intracardiac, intradermal,intraperitoneal, transtracheal, subcutaneous, subcuticular,intraarticular, subcapsular, subarachnoid, intraspinal, epidural andintrasternal injection and infusion, as well as in vivo electroporation.The TKI is typically administered via a non-parenteral route, preferablyorally. Other non-parenteral routes include a topical, epidermal ormucosal route of administration, for example, intranasally, vaginally,rectally, sublingually or topically. Administering can also beperformed, for example, once, a plurality of times, and/or over one ormore extended periods. Dose intervals denominated in “days” are intendedto represent approximately 24 hour intervals, but may vary slightly dueto scheduling difficulties or other delays in administration.

“Concurrent” administration refers to dosing of two distinct agents,such as anti-fucosyl GM1 and anti-CD137 antibodies, at or around thesame time rather than intentionally delaying administration of one ofthe agents. As such, concurrent administration includes simultaneousadministration, e.g. when the agents are co-formulated or mixed prior toadministration, and also includes administration of the two drugs withina convenient interval, typically during the same visit to a health carefacility. Typically concurrent administration is performed on the sameday, and excludes administration at separate visits to a health carefacility.

An “adverse event” (AE) as used herein is any unfavorable and generallyunintended or undesirable sign (including an abnormal laboratoryfinding), symptom, or disease associated with the use of a medicaltreatment. A medical treatment may have one or more associated AEs andeach AE may have the same or different level of severity. Reference tomethods capable of “altering adverse events” means a treatment regimethat decreases the incidence and/or severity of one or more AEsassociated with the use of a different treatment regime.

An “antibody” (Ab) shall include, without limitation, a glycoproteinimmunoglobulin that binds specifically to an antigen and comprises atleast two heavy (H) chains and two light (L) chains interconnected bydisulfide bonds, or an antigen-binding portion thereof. Each H chaincomprises a heavy chain variable region (abbreviated herein as V_(H))and a heavy chain constant region. The heavy chain constant regioncomprises three constant domains, C_(H1), C_(H2) and C_(H3). Each lightchain comprises a light chain variable region (abbreviated herein asV_(L)) and a light chain constant region. The light chain constantregion is comprises one constant domain, C_(L). The V_(H) and V_(L)regions can be further subdivided into regions of hypervariability,termed complementarity determining regions (CDRs), interspersed withregions that are more conserved, termed framework regions (FR). EachV_(H) and V_(L) comprises three CDRs and four FRs, arranged fromamino-terminus to carboxy-terminus in the following order: FR1, CDR1,FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and lightchains contain a binding domain that interacts with an antigen. Theconstant regions of the Abs may mediate the binding of theimmunoglobulin to host tissues or factors, including various cells ofthe immune system (e.g., effector cells) and the first component (C1q)of the classical complement system. Sequences of antibody heavy chainsherein may comprise a C-terminal lysine (K) residue but that residue maybe clipped off during manufacture, entirely or partially, or it may beremoved from the genetic construct used to produce the antibody so as toavoid potential heterogeneity arising from the aforementioned clipping.Both heavy chain sequences provided herein (SEQ ID NOs: 3 and 13) do notinclude C-terminal lysine residues.

An immunoglobulin may derive from any of the commonly known isotypes,including but not limited to IgA, secretory IgA, IgG and IgM. IgGsubclasses are also well known to those in the art and include but arenot limited to human IgG1, IgG2, IgG3 and IgG4. “Isotype” refers to theAb class or subclass (e.g., IgM or IgG1) that is encoded by the heavychain constant region genes. The term “antibody” includes, by way ofexample, both naturally occurring and non-naturally occurring Abs;monoclonal and polyclonal Abs; chimeric and humanized Abs; human ornonhuman Abs; wholly synthetic Abs; and single chain Abs. A nonhuman Abmay be humanized by recombinant methods to reduce its immunogenicity inman. Where not expressly stated, and unless the context indicatesotherwise, the term “antibody” also includes an antigen-binding fragmentor an antigen-binding portion of any of the aforementionedimmunoglobulins, and includes a monovalent and a divalent fragment orportion, and a single chain Ab.

An “isolated antibody” refers to an Ab that is substantially free ofother Abs having different antigenic specificities (e.g., an isolated Abthat binds specifically to fucosyl-GM1 is substantially free of Abs thatbind specifically to antigens other than fucosyl-GM1). Moreover, anisolated Ab may be substantially free of other cellular material and/orchemicals.

The term “monoclonal antibody” (“mAb”) refers to a non-naturallyoccurring preparation of Ab molecules of single molecular composition,i.e., Ab molecules whose primary sequences are essentially identical,and which exhibits a single binding specificity and affinity for aparticular epitope. A mAb is an example of an isolated Ab. MAbs may beproduced by hybridoma, recombinant, transgenic or other techniques knownto those skilled in the art.

A “human” antibody (HuMAb) refers to an Ab having variable regions inwhich both the framework and CDR regions are derived from human germlineimmunoglobulin sequences. Furthermore, if the Ab contains a constantregion, the constant region also is derived from human germlineimmunoglobulin sequences. The human Abs of the invention may includeamino acid residues not encoded by human germline immunoglobulinsequences (e.g., mutations introduced by random or site-specificmutagenesis in vitro or by somatic mutation in vivo). However, the term“human antibody,” as used herein, is not intended to include Abs inwhich CDR sequences derived from the germline of another mammalianspecies, such as a mouse, have been grafted onto human frameworksequences. The terms “human” Abs and “fully human” Abs and are usedsynonymously.

A “humanized antibody” refers to an Ab in which some, most or all of theamino acids outside the CDR domains of a non-human Ab are replaced withcorresponding amino acids derived from human immunoglobulins. In oneembodiment of a humanized form of an Ab, some, most or all of the aminoacids outside the CDR domains have been replaced with amino acids fromhuman immunoglobulins, whereas some, most or all amino acids within oneor more CDR regions are unchanged. Small additions, deletions,insertions, substitutions or modifications of amino acids arepermissible as long as they do not abrogate the ability of the Ab tobind to a particular antigen. A “humanized” Ab retains an antigenicspecificity similar to that of the original Ab.

A “chimeric antibody” refers to an Ab in which the variable regions arederived from one species and the constant regions are derived fromanother species, such as an Ab in which the variable regions are derivedfrom a mouse Ab and the constant regions are derived from a human Ab.

An “anti-antigen” Ab refers to an Ab that binds specifically to theantigen. For example, an anti-fucosyl-GM1 Ab binds specifically tofucosyl-GM1.

An “antigen-binding portion” of an Ab (also called an “antigen-bindingfragment”) refers to one or more fragments of an Ab that retain theability to bind specifically to the same antigen bound by the whole Ab.

A “cancer” refers a broad group of various diseases characterized by theuncontrolled growth of abnormal cells in the body. Unregulated celldivision and growth divide and grow results in the formation ofmalignant tumors that invade neighboring tissues and may alsometastasize to distant parts of the body through the lymphatic system orbloodstream. The terms, “cancer,” “tumor,” and “neoplasm,” are usedinterchangeably herein.

A “subject” includes any human or nonhuman animal. The term “nonhumananimal” includes, but is not limited to, vertebrates such as nonhumanprimates, sheep, dogs, and rodents such as mice, rats and guinea pigs.In preferred embodiments, the subject is a human. The terms, “subject”and “patient” are used interchangeably herein.

A “therapeutically effective amount” or “therapeutically effectivedosage” of a drug or therapeutic agent is any amount of the drug that,when used alone or in combination with another therapeutic agent,protects a subject against the onset of a disease or promotes diseaseregression evidenced by a decrease in severity of disease symptoms, anincrease in frequency and duration of disease symptom-free periods, or aprevention of impairment or disability due to the disease affliction.The ability of a therapeutic agent to promote disease regression can beevaluated using a variety of methods known to the skilled practitioner,such as in human subjects during clinical trials, in animal modelsystems predictive of efficacy in humans, or by assaying the activity ofthe agent in in vitro assays.

By way of example, an “anti-cancer agent” promotes cancer regression ina subject. In preferred embodiments, a therapeutically effective amountof the drug promotes cancer regression to the point of eliminating thecancer. “Promoting cancer regression” means that administering aneffective amount of the drug, alone or in combination with ananti-neoplastic agent, results in a reduction in tumor growth or size,necrosis of the tumor, a decrease in severity of at least one diseasesymptom, an increase in frequency and duration of disease symptom-freeperiods, or a prevention of impairment or disability due to the diseaseaffliction. In addition, the terms “effective” and “effectiveness” withregard to a treatment includes both pharmacological effectiveness andphysiological safety. Pharmacological effectiveness refers to theability of the drug to promote cancer regression in the patient.Physiological safety refers to the level of toxicity, or other adversephysiological effects at the cellular, organ and/or organism level(adverse effects) resulting from administration of the drug.

By way of example for the treatment of tumors, a therapeuticallyeffective amount of an anti-cancer agent preferably inhibits cell growthor tumor growth by at least about 20%, more preferably by at least about40%, even more preferably by at least about 60%, and still morepreferably by at least about 80% relative to untreated subjects. Inother preferred embodiments of the invention, tumor regression may beobserved and continue for a period of at least about 20 days, morepreferably at least about 40 days, or even more preferably at leastabout 60 days. Notwithstanding these ultimate measurements oftherapeutic effectiveness, evaluation of immunotherapeutic drugs mustalso make allowance for “immune-related” response patterns.

A therapeutically effective amount of a drug includes a“prophylactically effective amount,” which is any amount of the drugthat, when administered alone or in combination with an anti-neoplasticagent to a subject at risk of developing a cancer (e.g., a subjecthaving a pre-malignant condition) or of suffering a recurrence ofcancer, inhibits the development or recurrence of the cancer. Inpreferred embodiments, the prophylactically effective amount preventsthe development or recurrence of the cancer entirely. “Inhibiting” thedevelopment or recurrence of a cancer means either lessening thelikelihood of the cancer's development or recurrence, or preventing thedevelopment or recurrence of the cancer entirely.

The use of the alternative (e.g., “or”) should be understood to meaneither one, both, or any combination thereof of the alternatives. Asused herein, the indefinite articles “a” or “an” should be understood torefer to “one or more” of any recited or enumerated component.

The terms “about” or “comprising essentially of” refer to a value orcomposition that is within an acceptable error range for the particularvalue or composition as determined by one of ordinary skill in the art,which will depend in part on how the value or composition is measured ordetermined, i.e., the limitations of the measurement system. Forexample, “about” or “comprising essentially of” can mean within 1 ormore than 1 standard deviation per the practice in the art.Alternatively, “about” or “comprising essentially of” can mean a rangeof up to 20%. Furthermore, particularly with respect to biologicalsystems or processes, the terms can mean up to an order of magnitude orup to 5-fold of a value. When particular values or compositions areprovided in the application and claims, unless otherwise stated, themeaning of “about” or “comprising essentially of” should be assumed tobe within an acceptable error range for that particular value orcomposition.

As described herein, any concentration range, percentage range, ratiorange or integer range is to be understood to include the value of anyinteger within the recited range and, when appropriate, fractionsthereof (such as one tenth and one hundredth of an integer), unlessotherwise indicated. Such ranges further include the values as theboundaries of the ranges.

Various aspects of the invention are described in further detail in thefollowing subsections.

II. Anti-Fucosyl-GM1 Antibodies

HuMAbs that bind specifically to fucosyl-GM1 with high affinity havebeen disclosed in U.S. Pat. No. 8,383,118 and WO 2007/067992 (e.g.,human monoclonal antibodies 5B1, 5B1a, 7D4, 7E4, 13B8 and 18D5). Each ofthe HuMAbs disclosed in U.S. Pat. No. 8,383,118 has been demonstrated toexhibit one or more desirable functional properties: (1) specificallybinds to fucosyl-GM1; (2) binds to fucosyl-GM1 with high affinity (forexample with a K_(D) of 1×10⁻⁷ M or less); (c) binds to the human smallcell lung cancer cell line DMS-79 (Human SCLC ATCC #CRL-2049); and (d)inhibit growth of tumor cells in vitro or in vivo. Preferably, theantibody binds to fucosyl-GM1 with a K_(D) of 5×10⁻⁸ M or less, binds tofucosyl-GM1 with a K_(D) of 1×10⁻⁸ M or less, binds to fucosyl-GM1 witha K_(D) of 5×10⁻⁹ M or less, or binds to fucosyl-GM1 with a K_(D) ofbetween 1×10⁻⁸ M and 1×10⁻¹⁰ M or less. Standard assays to evaluate thebinding ability of the antibodies toward fucosyl-GM1 are known in theart, including for example, ELISAs, Western blots and RIAs. The bindingkinetics (e.g., binding affinity) of the antibodies also can be assessedby standard assays known in the art, such as by ELISA, Scatchard andBiacore analysis.

A preferred anti-fucosyl-GM1 Ab is BMS-986012 (also referred to asMDX-1110 or 7E4).

Anti-fucosyl-GM1 Abs usable in the disclosed methods also includeisolated Abs that bind specifically to fucosyl-GM1 and compete forbinding to fucosyl-GM1 with BMS-986012 (see, e.g., U.S. Pat. No.8,383,118; WO 2007/067992). The ability of Abs to compete for binding toan antigen indicates that these Abs bind to the same epitope region ofthe antigen and sterically hinder the binding of other competing Abs tothat particular epitope region. These competing Abs are expected to havefunctional properties very similar those of BMS-986012 by virtue oftheir binding to the same epitope region of fucosyl-GM1. Competing Abscan be readily identified based on their ability to compete withBMS-986012 in standard fucosyl-GM1 binding assays such as Biacoreanalysis, ELISA assays or flow cytometry (see, e.g., WO 2013/173223).

For administration to human subjects, these Abs are preferably chimericAbs, or more preferably humanized or human Abs. Such chimeric, humanizedor human mAbs can be prepared and isolated by methods well known in theart. Anti-fucosyl-GM1 Abs usable in the methods of the disclosedinvention also include antigen-binding portions of the above Abs. It hasbeen amply demonstrated that the antigen-binding function of an Ab canbe performed by fragments of a full-length Ab. Examples of bindingfragments encompassed within the term “antigen-binding portion” of an Abinclude (i) a Fab fragment, a monovalent fragment consisting of theV_(L), V_(H), C_(L) and C_(H1) domains; (ii) a F(ab′)₂ fragment, abivalent fragment comprising two Fab fragments linked by a disulfidebridge at the hinge region; (iii) a Fd fragment consisting of the V_(H)and C_(H)1 domains; and (iv) a Fv fragment consisting of the V_(L) andV_(H) domains of a single arm of an Ab. Anti-fucosyl-GM1 antibodies (orVH and/or VL domains derived therefrom) suitable for use in theinvention can be generated using methods well known in the art.

An exemplary anti-fucosyl-GM1 antibody is BMS-986012 comprising heavyand light chains comprising the sequences shown in SEQ ID NOs: 3 and 4,respectively, or antigen binding fragments and variants thereof.

In other embodiments, the antibody has heavy and light chain CDRs orvariable regions of BMS-986012. Accordingly, in one embodiment, theantibody comprises CDR1, CDR2, and CDR3 domains of the VH of BMS-986012having the sequence set forth in SEQ ID NO: 1, and CDR1, CDR2 and CDR3domains of the VL of BMS-986012 having the sequence set forth in SEQ IDNO: 2. In another embodiment, the antibody comprises the heavy chainCDR1, CDR2 and CDR3 domains comprising the sequences set forth in SEQ IDNOs: 5, 6, and 7, respectively, and the light chain CDR1, CDR2 and CDR3domains comprising the sequences set forth in SEQ ID NOs: 8, 9, and 10,respectively. In another embodiment, the antibody comprises VH and VLregions comprising the amino acid sequences set forth in SEQ ID NO: 1and SEQ ID NO: 2, respectively. In another embodiment, the antibodycompetes for binding with and/or binds to the same epitope onfucosyl-GM1 as the above-mentioned antibodies. In another embodiment,the antibody has at least about 90% variable region amino acid sequenceidentity with the above-mentioned antibodies (e.g., at least about 90%,95% or 99% variable region identity with SEQ ID NO: 1 or SEQ ID NO: 2).

II. Non-Fucosylation and Hypofucosylation

The interaction of antibodies with FcγRs can be enhanced by modifyingthe glycan moiety attached to each Fc fragment at the N297 residue. Inparticular, the absence of branching fucose residues strongly enhancesADCC via improved binding of IgG to activating FcγRIIIA without alteringantigen binding or CDC. Natsume et al. (2009) Drug Des. Devel. Ther.3:7. There is convincing evidence that afucosylated tumor-specificantibodies translate into enhanced therapeutic activity in mouse modelsin vivo. Nimmerjahn & Ravetch (2005) Science 310:1510; Mossner et al.(2010) Blood 115:4393.

Modification of antibody glycosylation can be accomplished by, forexample, expressing the antibody in a host cell with alteredglycosylation machinery. Antibodies with reduced or eliminatedfucosylation, which exhibit enhanced ADCC, are particularly useful inthe methods of the present invention. Cells with altered glycosylationmachinery have been described in the art and can be used as host cellsin which to express recombinant antibodies of this disclosure to therebyproduce an antibody with altered glycosylation. For example, the celllines Ms704, Ms705, and Ms709 lack the fucosyltransferase gene, FUT8(α-(1,6) fucosyltransferase (see U.S. Pat. App. Publication No.20040110704; Yamane-Ohnuki et al. (2004) Biotechnol. Bioeng. 87: 614),such that antibodies expressed in these cell lines lack fucose on theircarbohydrates. As another example, EP 1176195 also describes a cell linewith a functionally disrupted FUT8 gene as well as cell lines that havelittle or no activity for adding fucose to the N-acetylglucosamine thatbinds to the Fc region of the antibody, for example, the rat myelomacell line YB2/0 (ATCC CRL 1662). PCT Publication WO 03/035835 describesa variant CHO cell line, Lec13, with reduced ability to attach fucose toAsn(297)-linked carbohydrates, also resulting in hypofucosylation ofantibodies expressed in that host cell. See also Shields et al. (2002) JBiol. Chem. 277:26733. Antibodies with a modified glycosylation profilecan also be produced in chicken eggs, as described in PCT PublicationNo. WO 2006/089231. Alternatively, antibodies with a modifiedglycosylation profile can be produced in plant cells, such as Lemna. Seee.g. U.S. Publication No. 2012/0276086. PCT Publication No. WO 99/54342describes cell lines engineered to express glycoprotein-modifyingglycosyl transferases (e.g., beta(1,4)-N-acetylglucosaminyltransferaseIII (GnTIII)) such that antibodies expressed in the engineered celllines exhibit increased bisecting GlcNac structures which results inincreased ADCC activity of the antibodies. See also Umaña et al. (1999)Nat. Biotech. 17:176. Alternatively, the fucose residues of the antibodymay be cleaved off using a fucosidase enzyme. For example, the enzymealpha-L-fucosidase removes fucosyl residues from antibodies. Tarentinoet al. (1975) Biochem. 14:5516. Antibodies with reduced fucosylation mayalso be produced in cells harboring a recombinant gene encoding anenzyme that uses GDP-6-deoxy-D-lyxo-4-hexylose as a substrate, such asGDP-6-deoxy-D-lyxo-4-hexylose reductase (RMD), as described at U.S. Pat.No. 8,642,292. Alternatively, cells may be grown in medium containingfucose analogs that block the addition of fucose residues to theN-linked glycan or a glycoprotein, such as antibody, produced by cellsgrown in the medium. U.S. Pat. No. 8,163,551; WO 09/135181.

IV. Anti-CD137 Antibodies

HuMAbs that bind specifically to human CD137 with high affinity havebeen disclosed and claimed in U.S. Pat. No. 7,288,638 (human monoclonalantibodies human 4-1BB, also known as CD137). The sequences ofanti-huCD137 mAb BMS-663513 are provided herein at SEQ ID NOs: 11-20.

Anti-huCD137 antibody BMS-663513 is a powerful CD137 agonist drivingimmune response and as a consequence must be administered at arelatively low dose to avoid immune-mediated side effects. In specificembodiments of monotherapy using BMS-663513, the antibody isadministered at a flat doses between 1 mg and 10 mg, e.g. 3 mg or 8 mg,at intervals of every 4 or 8 weeks. See WO 2016/029073. In otherembodiments BMS-663513 can be administered at between 0.05 and 1 mg/kg,e.g. 0.1 or 0.3 mg/kg.

Anti-huCD137 Abs usable in the disclosed methods also include isolatedAbs that bind specifically to CD137 and compete for binding to CD137with BMS-663513. The ability of Abs to compete for binding to an antigenindicates that these Abs bind to the same epitope region of the antigenand sterically hinder the binding of other competing Abs to thatparticular epitope region. These competing Abs are expected to havefunctional properties very similar those of BMS-663513 by virtue oftheir binding to the same epitope region of CD137. Competing Abs can bereadily identified based on their ability to compete with BMS-663513 instandard binding assays such as Biacore analysis, ELISA assays or flowcytometry.

For administration to human subjects, these Abs are preferably chimericAbs, or more preferably humanized or human Abs. Such chimeric, humanizedor human mAbs can be prepared and isolated by methods well known in theart. Anti-huCD137 Abs usable in the methods of the disclosed inventionalso include antigen-binding portions of the above Abs. Examples ofbinding fragments encompassed within the term “antigen-binding portion”of an Ab include (i) a Fab fragment, a monovalent fragment consisting ofthe V_(L), V_(H), C_(L) and C_(H1) domains; (ii) a F(ab′)₂ fragment, abivalent fragment comprising two Fab fragments linked by a disulfidebridge at the hinge region; (iii) a Fd fragment consisting of the V_(H)and C_(H1) domains; and (iv) a Fv fragment consisting of the V_(L) andV_(H) domains of a single arm of an Ab.

V. Antibodies that Compete with Reference Antibodies

Antibodies that compete with anti-fucosyl GM1 mAb BMS-986012 oranti-CD137 mAb BMS-663513 and for binding to their respective antigensmay be determined using methods well known in the art, for example in anenzyme linked immunosorbent assay (ELISA) or by fluorescence-activatedcell sorting (FACS).

An exemplary competition experiment to determine whether a test antibody“competes with” a reference antibody (BMS-986012 or BMS-663513) may beconducted as follows: cells expressing the antigen (fucosyl-GM1 orCD137) are seeded at 10⁵ cells per sample well in a 96 well plate. Theplate is set on ice followed by the addition of unconjugated testantibody at concentrations ranging from 0 to 50 pg/mL (three-foldtitration starting from a highest concentration of 50 μg/mL). Anunrelated IgG may be used as an isotype control for the first antibodyand added at the same concentrations (three-fold titration starting froma highest concentration of 50 μg/mL). A sample pre-incubated with 50μg/mL unlabeled reference antibody may be included as a positive controlfor complete blocking (100% inhibition) and a sample without antibody inthe primary incubation may be used as a negative control (nocompetition; 0% inhibition). After 30 minutes of incubation, labeled,e.g., biotinylated, reference antibody is added at a concentration of 2μg/mL per well without washing. Samples are incubated for another 30minutes on ice. Unbound antibodies are removed by washing the cells withFACS buffer. Cell-bound labeled reference antibody is detected with anagent that detects the label, e.g., PE conjugated streptavidin(Invitrogen, catalog #521388) for detecting biotin. The samples areacquired on a FACS Calibur Flow Cytometer (BD, San Jose) and analyzedwith Flowjo software (Tree Star, Inc., Ashland, Oreg.). The results maybe represented as % inhibition.

Unless otherwise indicated, an antibody will be considered to competewith a reference antibody (BMS-986012 or BMS-663513) if it reducesbinding of the selected antibody to its respective antigen by at least20% when used at a roughly equal molar concentration with the referenceantibody, as measured in a competition ELISA as outlined in thepreceding paragraph.

VI. Pharmaceutical Compositions

Therapeutic agents (e.g., anti-fucosyl-GM1 antibodies and/or anti-CD137antibodies, or antigen binding fragments thereof) of the presentinvention may be constituted in a composition, e.g., a pharmaceuticalcomposition containing and a pharmaceutically acceptable carrier.Pharmaceutical compositions of the present invention include bothindividual antibodies and co-formulations.

As used herein, a “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like that arephysiologically compatible. “Pharmaceutically acceptable” means approvedby a government regulatory agency or listed in the U.S. Pharmacopeia oranother generally recognized pharmacopeia for use in animals,particularly in humans. The term “carrier” refers to a diluent,adjuvant, excipient, or vehicle with which the compound is administered.Such pharmaceutical carriers can be sterile liquids, such as water andoils, including those of petroleum, animal, vegetable or syntheticorigin, such as peanut oil, soybean oil, mineral oil, sesame oil,glycerol polyethylene glycol ricinoleate, and the like. Water or aqueoussolution saline and aqueous dextrose and glycerol solutions may beemployed as carriers, particularly for injectable solutions (e.g.,comprising an anti-fucosyl-GM1 antibody). Preferably, the carrier for acomposition containing an Ab is suitable for intravenous, intramuscular,subcutaneous, parenteral, spinal or epidermal administration (e.g., byinjection or infusion). A pharmaceutical composition of the inventionmay include one or more pharmaceutically acceptable salts, anti-oxidant,aqueous and non-aqueous carriers, and/or adjuvants such aspreservatives, wetting agents, emulsifying agents and dispersing agents.

Liquid compositions for parenteral administration can be formulated foradministration by injection or continuous infusion. Routes ofadministration by injection or infusion include intravenous,intraperitoneal, intramuscular, intrathecal and subcutaneous. In oneembodiment, the anti-fucosyl-GM1 antibody is administered intravenously.

VII. Methods of Treatment

Provided herein are clinical methods for treating a lung cancer (e.g.,small cell lung cancer) in a subject (e.g., a human subject), comprisingadministering to the subject a therapeutically effective amounts of ananti-fucosyl-GM1 antibody, or an antigen-binding portion thereof and ananti-huCD137 antibody, or an antigen-binding portion thereof. In oneembodiment, the subject has previously received an initial anti-cancertherapy. In another embodiment, the lung cancer is an advanced,metastatic, relapsed, and/or refractory lung cancer.

In a particular embodiment, the anti-fucosyl-GM1 antibody is BMS-986012.In some embodiments, the anti-huCD137 antibody is BMS-663513. In anotherembodiment, dosage regimens are adjusted to provide the optimum desiredresponse (e.g., an effective response).

In another embodiment, the antibody is administered as a first line oftreatment (e.g., the initial or first treatment). In another embodiment,the antibody is administered as a second line of treatment (e.g., afterthe initial or first treatment, including after relapse and/or where thefirst treatment has failed).

In certain embodiments, the anti-fucosyl-GM1 antibody is administered ata dose ranging from 10 to 2000 mg once every 1, 2, 3 or 4 weeks. Forexample, the antibody is administered at a dose ranging from 20 to 1000mg once every 3 weeks. Optionally, the method comprises at least onetreatment cycle of three weeks. For example, the method comprises atleast four treatment cycles of three weeks. To illustrate, the antibodyis administered on Days 1, 22, 43, and 64.

In certain specific embodiments, the antibody is administered accordingto at least one of the following dosing regimens: (a) about 20 mg of theantibody every 3 weeks; (b) about 70 mg of the antibody every 3 weeks;(c) about 160 mg of the antibody every 3 weeks; (d) about 400 mg of theantibody every 3 weeks; and (e) about 1000 mg of the antibody every 3weeks. In one embodiment antibody is administered at a dose between 400and 1000 mg, inclusive. Whether stated or not, any dose range recitedherein is intended to be inclusive, i.e. it the doses recited as theboundaries of the ranges are included within the recited dosing range.Preferably, administration of the antibody induces a durable clinicalresponse in the subject. Optionally, administration of the antibody iscontinued for as long as clinical benefit is observed or untilunmanageable toxicity or disease progression occurs. The efficacy of thetreatment methods provided herein can be assessed using any suitablemeans. In one embodiment, the treatment produces at least onetherapeutic effect selected from the group consisting of reduction insize of the cancer, reduction in number of metastatic lesions over time,complete response, partial response, and stable disease.

Patients treated according to the methods disclosed herein preferablyexperience improvement in at least one sign of cancer. In oneembodiment, improvement is measured by a reduction in the quantityand/or size of measurable tumor lesions. In another embodiment, lesionscan be measured on chest x-rays or CT or MRI films. In anotherembodiment, cytology or histology can be used to evaluate responsivenessto a therapy.

In one embodiment, the patient treated exhibits a complete response(CR), a partial response (PR), or stable disease (SD). In anotherembodiment, the patient treated experiences tumor shrinkage and/ordecrease in growth rate, i.e., suppression of tumor growth. In anotherembodiment, unwanted cell proliferation is reduced or inhibited. In yetanother embodiment, one or more of the following can occur: the numberof cancer cells can be reduced; tumor size can be reduced; cancer cellinfiltration into peripheral organs can be inhibited, retarded, slowed,or stopped; tumor metastasis can be slowed or inhibited; tumor growthcan be inhibited; recurrence of tumor can be prevented or delayed; oneor more of the symptoms associated with cancer can be relieved to someextent.

VIII. Kits and Unit Dosage Forms

Also provided herein are kits that include a pharmaceutical compositioncontaining an anti-fucosyl-GM1 antibody (such as BMS-986012) and/or ananti-huCD137 antibody (BMS-663513), and a pharmaceutically acceptablecarrier, in a therapeutically effective amount adapted for use in thepreceding methods.

The kits optionally can also include instructions, e.g., comprisingadministration schedules, to allow a practitioner (e.g., a physician,nurse, or patient) to administer the composition contained therein toadminister the composition to a patient having a cancer (e.g., a lungcancer). The kit can also include a syringe.

Optionally, the kits include multiple packages of the single-dosepharmaceutical compositions each containing an effective amount of theantibody for a single administration in accordance with the methodsprovided above. Instruments or devices necessary for administering thepharmaceutical composition(s) also may be included in the kits. Forinstance, a kit may provide one or more pre-filled syringes containingan amount of the antibody.

The following examples are merely illustrative and should not beconstrued as limiting the scope of this disclosure in any way as manyvariations and equivalents will become apparent to those skilled in theart upon reading the present disclosure.

The contents of all references, GenBank entries, patents and publishedpatent applications cited throughout this application are expresslyincorporated herein by reference.

Example 1 Combination Therapy-Anti-Fucosyl GM1 &Anti-CD137

A combination therapy method of the present invention involvingantibodies to fucosyl GM1 and CD137 was tested in the DMS-79 (ATCCCRL-2049) SCLC tumor model in mice. Bepler et al. (1989) Oncogene 4:45;Pettengill (1980) Cancer 45: 906; Pettengill et al. (1980) Exp. CellBiol. 48:279. Briefly, DMS79 cells were cultured in RPMI with 10% fetalbovine serum (FBS), 2 mM L-glutamine, 15 mg/L sodium bicarbonate, 4.5g/L glucose, 10 mM HEPES, and 10 μM NaPyr prior to subcutaneousimplantation in the right flanks of male C.B17 SCID mice (5 millioncells in 0.1 mL phosphate-buffered saline (PBS) and 0.1 mL MATRIGEL®gelatinous protein mixture per flank). When tumors reached a mean ormedian size of 80-155 mm³ estimated by L×W×H/2 using a digital calipers,mice were randomized into treatment groups (N=8 mice per group).

Anti-fucosyl GM1 antibody BMS-986012 was administered at 3 mg/kg i.p. tomice on days 6, 10, 13, 17 and 20 post-implantation to assess itseffectiveness as monotherapy. Because fucosyl GM1 is a gangliosiderather than a protein, and is the same in mice as in humans, there wasno need to use a “mouse surrogate” for mouse studies. Anti-muCD137antibody (a mouse surrogate for BMS-663513) was administered to mice at1 mg/kg i.p. on days 7, 11, 14, 18 and 21 post implantation to assessits effectiveness as monotherapy.

Combination therapy involved administration of both antibodies accordingto the same schedule, i.e. anti-CD137 (either 0.1 or 0.3 mg/kg) wasadministered one day after each dose of anti-fucosyl GM1 (3 mg/kg). Avehicle only control was included. Results are provided at FIG. 1.Anti-CD137 monotherapy was ineffective, while anti-fucosyl GM1monotherapy significantly reduced tumor growth. Combination therapy, incontrast, virtually eliminated tumor growth at both doses of anti-CD137tested, demonstrating the synergistic effect of the two antibodies.

Example 2 Combination Therapy—Anti-Fucosyl GM1 & Cisplatin

A combination therapy method of the present invention involving antibodyto fucosyl GM1 and cisplatin was tested in the DMS-79 SCLC tumor modelin mice (N=9 mice per group), essentially as described in Example 1.Anti-fucosyl GM1 antibody BMS-986012 was administered at 0.3 mg/kg i.p.to mice on days 7, 10, 13, 17 and 21 post implantation, and cisplatinwas administered at 3 mg/kg on days 7, 14, 21 and 28. Cisplatin wasadministered in combination with BMS-986012 and with an isotype controlantibody at 3 mg/kg as a control. Additional controls included theisotype control antibody alone and a vehicle control. Results areprovided at FIG. 2. While both anti-fucosyl GM1 antibody treatment andcisplatin treatment are effective as monotherapy to reduce tumor growth,the combination is significantly more effective—almost stopping tumorgrowth entirely.

Example 3 Combination Therapy—Anti-Fucosyl GM1 & Etoposide

A combination therapy method of the present invention involving antibodyto fucosyl GM1 and etoposide was tested in the DMS-79 SCLC tumor modelin mice (N=9 mice per group), essentially as described in Example 1.Anti-fucosyl GM1 antibody BMS-986012 was administered at 3 mg/kg i.p. tomice on days 7, 11, 15, 18 and 21 post-implantation, and etoposide wasadministered at 15 mg/kg i.p. on days 7, 9 and 11. Etoposide wasadministered in combination with BMS-986012 and with an isotype controlantibody at 3 mg/kg as a control. Additional controls included theisotype control antibody alone and a vehicle control. Results areprovided at FIG. 3. While both anti-fucosyl GM1 antibody treatment andetoposide treatment are somewhat effective as monotherapy to reducetumor growth, the combination is more effective.

TABLE 1 SUMMARY OF SEQUENCE LISTING SEQ ID NO. Description  1Anti-FucGM1 Heavy Chain Variable Domain  2 Anti-FucGM1 Light ChainVariable Domain  3 Anti-FucGM1 Heavy Chain  4 Anti-FucGM1 Light Chain  5Anti-FucGM1 CDRH1  6 Anti-FucGM1 CDRH2  7 Anti-FucGM1 CDRH3  8Anti-FucGM1 CDRL1  9 Anti-FucGM1 CDRL2 10 Anti-FucGM1 CDRL3 11Anti-CD137 Heavy Chain Variable Domain 12 Anti-CD137 Light ChainVariable Domain 13 Anti-CD137 Heavy Chain 14 Anti-CD137 Light Chain 15Anti-CD137 CDRH1 16 Anti-CD137 CDRH2 17 Anti-CD137 CDRH3 18 Anti-CD137CDRL1 19 Anti-CD137 CDRL2 20 Anti-CD137 CDRL3 21 huCD137-NP_001552

Antibody sequences in the Sequence Listing include the sequences of themature variable regions of the heavy and light chains, i.e. thesequences do not include signal peptides.

Equivalents: Those skilled in the art will recognize, or be able toascertain using no more than routine experimentation, many equivalentsof the specific embodiments disclosed herein. Such equivalents areintended to be encompassed by the following claims.

What is claimed is:
 1. A method for treating a subject afflicted withcancer, comprising administering to the subject a therapeuticallyeffective combination of: a. an anti-fucosyl GM1 antibody, or anantigen-binding portion thereof; and b. an anti-CD137 antibody, or anantigen-binding portion thereof.
 2. The method of claim 1, wherein theanti-fucosyl GM1 antibody is hypofucosylated or non-fucosylated.
 3. Themethod of claim 2, wherein the cancer is lung cancer.
 4. The method ofclaim 3, wherein the lung cancer is small cell lung cancer (SCLC). 5.The method of claim 1, wherein the anti-fucosyl GM1 antibody competeswith BMS-986012 for binding to fucosyl-GM1, and further whereinBMS-986012 comprises heavy and light chains comprising the sequences ofSEQ ID NOs: 3 and 4, respectively.
 6. The method of claim 5, wherein theanti-fucosyl GM1 antibody, or antigen-binding fragment thereof,comprises: a. a heavy chain comprising: i. CDRH1 comprising the sequenceof SEQ ID NO: 5; ii. CDRH2 comprising the sequence of SEQ ID NO: 6; andiii. CDRH3 comprising the sequence of SEQ ID NO: 7; and b. a light chaincomprising: i. CDRL1 comprising the sequence of SEQ ID NO: 8; ii. CDRL2comprising the sequence of SEQ ID NO: 9; and iii. CDRL3 comprising thesequence of SEQ ID NO:
 10. 7. The method of claim 6, wherein theanti-fucosyl GM1 antibody, or antigen-binding fragment thereof,comprises: a. a heavy chain variable region comprising the sequence ofSEQ ID NO: 1; and b. a light chain variable region comprising thesequence of SEQ ID NO:
 2. 8. The method of claim 7, wherein theanti-fucosyl GM1 antibody is non-fucosylated.
 9. The method of claim 8,wherein the anti-fucosyl GM1 antibody is BMS-986012, wherein BMS-986012comprises heavy and light chains comprising the sequences of SEQ ID NOs:3 and 4, respectively.
 10. The method of claim 7, wherein theanti-fucosyl GM1 antibody is an antibody-drug conjugate.
 11. The methodof claim 1, wherein the anti-huCD137 antibody competes with BMS-663513for binding to human CD137, and further wherein BMS-663513 comprisesheavy and light chains comprising the sequences of SEQ ID NOs: 13 and14, respectively.
 12. The method of claim 11, wherein the anti-huCD137antibody, or antigen-binding fragment thereof, comprises: a. a heavychain comprising: i. CDRH1 comprising the sequence of SEQ ID NO: 15; ii.CDRH2 comprising the sequence of SEQ ID NO: 16; and iii. CDRH3comprising the sequence of SEQ ID NO: 17; and b. a light chaincomprising: i. CDRL1 comprising the sequence of SEQ ID NO: 18; ii. CDRL2comprising the sequence of SEQ ID NO: 19; and iii. CDRL3 comprising thesequence of SEQ ID NO:
 20. 13. The method of claim 12, wherein theanti-huCD137 antibody, or antigen-binding fragment thereof, comprises:a. a heavy chain variable region comprising the sequence of SEQ ID NO:11; and b. a light chain variable region comprising the sequence of SEQID NO:
 12. 14. The method of claim 13, wherein the anti-huCD137 antibodyis BMS-663513, wherein BMS-663513 comprises heavy and light chainscomprising the sequences set forth in SEQ ID NOs: 13 and 14,respectively.
 15. The method of claim 14, wherein the anti-fucosyl GM1antibody is BMS-986012, wherein BMS-986012 comprises heavy and lightchains comprising the sequences of SEQ ID NOs: 3 and 4, respectively.16. The method of claim 1, wherein the anti-fucosyl GM1 antibody, orantigen-binding portion thereof, is administered at a dose between 400and 1000 mg.
 17. The method of claim 1, wherein the anti-huCD137 GM1antibody, or antigen-binding portion thereof, is administered at a dosebetween 3 and 8 mg.
 18. The method of claim 1, wherein the anti-fucosylGM1 antibody, or antigen-binding portion thereof, is administeredconcurrently with administration of the anti-CD137 antibody, orantigen-binding portion thereof.
 19. The method of claim 1, wherein theanti-fucosyl GM1 antibody, or antigen-binding portion thereof, isadministered prior to administration of the anti-CD137 antibody, orantigen-binding portion thereof.
 20. The method of claim 19 wherein theanti-fucosyl GM1 antibody, or antigen-binding portion thereof, isadministered approximately one day prior to administration of theanti-CD137 antibody, or antigen-binding portion thereof.